Rhodococcus erythropolis and its uses in crude oil desulfuration
A technology of Rhodococcus erythropolis and crude oil, applied in bacteria and other directions, can solve problems such as increasing desulfurization cost, and achieve the effect of great application prospects
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Embodiment 1
[0049] Example 1: Screening of Rhodococcus erythropolis CCTCC M205068 bacterial strain
[0050] Various contaminated soils near oil fields or refineries were collected. Each weighed 5 grams of the above soil was added to 50 mL of liquid inorganic salt basic medium containing 0.01% dibenzothiophene by mass volume ratio, and cultured with shaking at 30 ° C for 48 hours at a speed of 250 rpm, and then 10 Inoculate the inoculation amount of % volume ratio into a new liquid inorganic salt basal medium containing 0.01% dibenzothiophene by mass volume ratio, at 30°C, under the condition of rotating speed 250 rpm, shake culture for 48 hours, and then continue Transfer, repeat this process 3 to 5 times. Dilute the cultured bacterial solution 1,000,000 times and spread it on the solid inorganic salt basic medium containing agar with a mass volume ratio of 1.6% and adding dibenzothiophene with a mass volume ratio of 0.01%, and culture it statically at 30°C for 72 hours . Select the gr...
Embodiment 2
[0056] Embodiment 2: Extraction of the 16SrDNA gene of the above-mentioned Rhodococcus erythropolis CCTCC M205068 bacterial strain
[0057] Cultivate 5ml of bacterial culture to saturation, take 1.5ml of culture, and centrifuge at 6000 rpm for 2 minutes; add 565μl of TE buffer to the precipitate, and the formula of TE buffer is as follows: 10mmol / L Tris (Tris), 1mmol / L ethylenediaminetetraacetic acid (EDTA), adjust the pH to 8.0 with hydrochloric acid, resuspend it by blowing repeatedly with a pipette, and add 30 μl of 10% sodium dodecylsulfonate ( SDS) and 5μl 20mg / mL proteinase K, mix well, and incubate at 37°C for 1 hour; add 100μl, 5mol / L NaCl, mix well, then add 80μl CTAB / NaCl solution, the CTAB / NaCl solution formula As follows: cetyltriethylammonium bromide (CTAB) with a mass volume ratio of 10% was dissolved in 0.7mol / L NaCl, mixed evenly, and incubated at 65°C for 10 minutes; an equal volume of chloroform / iso Pentanol, mix well, centrifuge at 12,000 rpm for 5 minutes,...
Embodiment 3
[0083] Embodiment 3: The method for removing organic sulfur in crude oil by using resting cells of Rhodococcus erythroflates, wherein the volume ratio of added crude oil to water phase is 1:10, and the treatment temperature is 25°C
[0084] The present invention utilizes the application of Rhodococcus erythropolis in crude oil desulfurization, and the sequence of steps involved in its desulfurization method is as follows:
[0085] (1) Strain selection: choose Rhodococcus erythropolis CCTCC M205068;
[0086] (2) Slant culture: Inoculate the bacteria on a solid slant inorganic salt basic medium containing 1.6% agar with a mass volume ratio of 0.005% dibenzothiophene, at 25° C. Cultivate for 42 hours;
[0087](3) Seed culture: the bacterial strain that step (2) is cultivated, connects 2 rings with inoculation loop under aseptic condition in 100mL and contains the liquid inorganic salt basic medium that mass volume ratio is 0.005% dibenzothiophene, 25 Under the condition of ℃, s...
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